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In vitro culture and characterization of duck primordial germ cells

机译:鸭原子胚芽细胞的体外培养与表征

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摘要

This study aimed to isolate, culture, and characterize duck primordial germ cells (PGCs) and to compare these cells with chicken PGCs. We first cultured Muscovy duck (Cairina moschata) circulating PGCs and gonadal PGCs (gPGCs) in the modified serum-containing medium used to amplify chicken PGCs. gPGCs were found to proliferate better in serum-free chemically defined medium than in serum-containing medium. Thereafter, gPGCs were similarly isolated from 2 other duck breeds, the Pekin duck (Anas platyrhynchos) and the hybrid mule duck (C. moschata x A. platyrhynchos), and amplified for a limited period of time in the chemically defined culture condition, but sufficiently to be characterized and transplanted. Cultured gPGCs of all 3 duck breeds were characterized by Periodic acid-Schiff staining, immunocytochemical staining, and expression analysis of germline-specific and pluripotency genes. Cultured duck gPGCs colonized the gonads after being genetically labeled and injected into recipient embryos. Taken together, these results demonstrate that duck PGCs retain their germline characteristics after being isolated, expanded in vitro, and genetically modified. Further studies are required to establish the optimal conditions for long-term culture of duck PGCs, which may involve supplementing the culture medium with other growth factors or compounds.
机译:本研究旨在分离,培养和表征鸭原族生殖细胞(PGCs)并将这些细胞与鸡PGC进行比较。我们在用于扩增鸡PGC的改性血清培养基中,我们首先培养的葡萄糖鸭(Cairina MoSchata)循环PGC和Gonadal PGC(GPGCS)。发现GPGC在无含血清培养基中的血清化学定义的培养基中更好地增殖。此后,GPGC类似地分离出2种其他鸭蛋白,皮鸭(ANAS platyrhynchos)和杂交骡鸭(C.Moschata X A.Platyrhynchos),并在化学定义的培养条件下在有限的时间内扩增,但是充分地表征和移植。通过定期酸性 - 席夫染色,免疫细胞化学染色和种系特异性和多能性基因的表达分析,表征了所有3只鸭品种的培养的GPGC。培养的鸭子GPGC在遗传标记并注射到受体胚胎后殖民化。总之,这些结果表明,鸭PGC在隔离后保留其种系特征,在体外扩增和遗传修饰。需要进一步的研究来建立鸭PGC的长期培养的最佳条件,这可能涉及补充培养基与其他生长因子或化合物。

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